BIODIVERSITAS ISSN: 1412-033X Volume 20, Number 1, January 2019 E-ISSN: 2085-4722 Pages: 54-60 DOI: 10.13057/biodiv/d200107

Taxonomy confirmation of cf. binotatus from Gunung Tujuh Lake, Jambi, Indonesia based on Cytochrome Oxidase-I (COI) gene

DEWI IMELDA ROESMA♥, DJONG HON TJONG, WILA KARLINA, DYTA RABBANI AIDIL Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Andalas. Jl. Universitas Andalas, Limau Manih, Pauh, Padang 25163, West Sumatra, Indonesia. Tel. +62-751- 777427. Fax. +62-751-71343, ♥email: [email protected].

Manuscript received: 21 September 2018. Revision accepted: 2 December 2018.

Abstract. Roesma D. I, Tjong D. H, Karlina W, Aidil D. R. 2019. confirmation of Puntius cf. binotatus from Gunung Tujuh Lake based on Cytochrome Oxidase-I (COI) gene. Biodiversitas 20: 54-60. The population of Puntius cf. binotatus Gunung Tujuh Lake in considered as an isolated population which found in the 20-30 meters depth under the water Gunung Tujuh Lake, Sumatra Island. The often exhibit different phenotypic characters and may genetic characters may also different due to their response to habitat/ecological conditions. Previous study on P. binotatus from several locations in West Sumatra showed the complexity of the genetic and morphological characters. A molecular study using COI gene of P. cf. binotatus from Gunung Tujuh Lake in Sumatera has been conducted to determine the taxonomic status of the . The distinct morphological characters of Puntius cf. binotatus is one black spot in the middle of the caudal and the absent of black spot on the base of the anterior dorsal ray. The analysis on COI gene showed that the range of sequence divergences between P. cf. binotatus Gunung Tujuh Lake and other Sumatra P. cf. binotatus is 3.1-7.6% and 4.1% to banksi. This value represents the differences at the subspecies level of Barbodes banksi. Therefore Barbodes banksi gunungtujuh is proposed as an appropriate name to P. cf. binotatus Gunung Tujuh Lake.

Keywords: Morphology, Cyt b gene, COI gene, Gunung Tujuh Lake, Puntius cf. binotatus

INTRODUCTION consisting of five rivers (Salibutan, Batang Kuranji, Sungai Asam, Batang Gumanti and Batang Sinamar) and one lake Gunung Tujuh Lake, Kerinci District, Jambi Province is (Lake Maninjau) in West Sumatra showed significant located in Sumatra Island, Indonesia at an altitude of 1,950 morphological differences among the locations. Recent meters above sea level, surrounded by volcanic mountains study, Karlina et al. (2016) using cyt b gene on P. cf. that have seven peaks. There is no river flow into the lake binotatus from Gunung Tujuh Lake and P. cf. Binotatus as the water source for the lake, except for water catchment recommended that P. cf. binotatus from Gunung Tujuh surrounding that entering the lake through the small rivers. Lake as a candidate of a new species. Water flows out of the lake directly forming a 75 meters According to Kottelat (2013), Puntius and Barbodes are high waterfall. It is believed that the fish live in this lake synonymous. Previously Taki et al. (1978), Mohsin and have been isolated for a long period. It has been proven that Ambak (1983), and Rainboth (1996) used the , the population in perfectly isolated habitats that prevent the , Barbodes, , , and Hypsibarbus as migration of individuals between populations. If there is the generic names. Roesma et al. (2018) reported those limited gene flow between populations, the frequency of genera are not synonym and P. cf. binotatus is suggested as genes in a population will be different from before, in Barbodes banksi from West Sumatra that consist of at least which the genetic mixing between populations still occur three subspecies based on Cytochrome Oxidase-I (COI) (Roesma et al. 2017). Handika et al. (2014) reported that gene. Based on the previous studies, it is crucial to use there is a type of fish with Puntius in Gunung Tujuh CO1 gene to identify and verify the taxonomic status of Lake that live in the 20-30 meters depth under the lake. Silver (P. cf. binotatus) from Gunung Tujuh Lake. A Based on the phenotypic characters identified, the species mtDNA gene has been widely used for taxonomic, is similar to P. binotatus, so it is identified as P. cf. phylogenetic and phylogeography studies in the Silver binotatus. According to Champasri et al. (2007) and Barb (John et al. 2013; Singh et al. 2013; Negi et al. 2017). Collins et al. (2012), Silver Barb or Spotted Barb are the The COI gene is a part of mtDNA used for species common names of Puntius, and they are sold for food and identification (DNA barcode). The DNA barcode is ornamental fish. designed to develop a rapid species identification that also Based on molecular studies using the cytochrome b allows detecting new species (Hebert et al. 2003a; Stoeckle gene (cyt b) mitochondrial DNA, Roesma (2011) and 2003; Hebert and Gregory 2005). COI gene has rapid Roesma et al. (2016) reported that P. binotatus from evolutionary rates than other mtDNA genes. Therefore, is several locations in West Sumatra reflected the complex useful for identification at species level, and has been and variable genetic and morphological characters among applied to identify numerous taxa (Hebert et al. the locations and considered them as P. cf. binotatus. Vitri 2003a; Clare et al. 2006; Hajibabaei et al. 2007; Smith et et al. (2012) reported that P. binotatus from six populations al. 2008; Frezal and Leblois 2008). ROESMA et al. – Taxonomy confirmation of Puntius cf. binotatus 55

MATERIAL AND METHODS 30 seconds, elongation at 72ºC for 1 minute and a final extension at 72ºC for 10 minutes. The PCR products were A total of 50 individuals of P. cf. binotatus in Gunung visualized in 2% agarose at 100 Volt, 20 W for 50 min. Tujuh Lake, Kerinci District, Jambi Province, Indonesia Good quality PCR product was used as samples for were collected following Cailiet et al. (1986). A piece of sequencing, performed in MacroGen USA DNA the tissue samples collected from individuals and stored in Sequencing Laboratory. Eppendorf tubes (1.5 mL) containing 96% ethanol PA. A total of 46 sequences of COI gene were analyzed Individual samples were temporarily stored in 10% consisted of four sequences Puntius cf. binotatus in formalin and transferred in 70% ethanol for long-term Gunung Tujuh Lake, 17 sequences P. cf. binotatus of other storage. Morphological measurements were done for 29 populations in Sumatra and 25 sequences taken from morphometric and 17 meristic characters following Genbank NCBI included outgroup species (Rasbora Kottelat et al. (1993) and Haryono and Tjakrawidjaja daniconius, Danio choprai, and rostrata) (Table 1). (2006) with additional three characters: Lenght of upper DNA sequences (forward and reverse) were assembled and caudal (LUCL), Lenght of lower caudal (LLCL) and edited using the DNA STAR program (Burland 2000). Lenght of middle caudal (LMCL). Morphometric and DNA sequence of P. cf. binotatus compared with the meristic characters were measured using digital calipers to sequences contained in NCBI, to check the similarity of all the nearest 0.1 mm. sequences in http: //blast.ncbi.nlm.nih.gov/Blast.cgi. All Total genomic DNA was extracted using Invitrogen sequences were aligned using the Clustal X program PureLinkTM Genomic DNA Mini Kit. The result of DNA (Thompson 1997). The aligned sequences were edited extraction was used for Polymerase chain reaction (PCR) using BIOEDIT program (Hall 1999). The number of amplification using thermal cycler (SensoQuest). PCR was polymorphic sequences was analyzed using the DNA conducted in 25 μl volumes containing 2x MyTaq Hs Red Sequence Polymorphism 5.10 (Rozas 2003). Sequence Mix (Bioline), 0.01 mM forward and reverse primer, 35 divergences of all sequences were analyzed using the ng/μl DNA template and 10 μl nuclear-free water. Primer Kimura-2 parameter model (K2P) in the MEGA 6.0 used were forward (Fish F1: 5'TCAACCAACCACAAA program (Tamura 2013). The phylogenetic tree was GACATTGG CAC3') and reverse (Fish R1: reconstructed based on four methods; Neighbors Joining 5'TAGACTTCTGGGTGGCCAAAGAATCA3') (Ward et (NJ), Maximum Evolution (ME), Maximum Likelihood al. 2005). The optimal PCR condition consisting of initial (ML) and Maximum Parsimony (MP) using MEGA 6.0 denaturation at 95°C for 2 minutes, 35 cycles consisting of (Tamura 2013). denaturation at 94ºC for 30 seconds, annealing at 54ºC for

Table 1. List of freshwater fish species barcoded along with GenBank Accession Numbers

Family Genus Species Location Accession No. Sources Puntius Puntius binotatus Lampung JQ665834 Wibowo et al. (2012) Puntius binotatus Malaysia JN646096 Sade and Biun (2012) Puntius banksi Malaysia JF781235 Song et al. (2013) Puntius sophore India JX983465 Khedkar et al. (2014) Puntius denisonii India HQ176003 Lakra et al. (2010) Puntius orphoides JF915642 Collins et al. (2011) Puntius sarana India EU417805 Lakra et al. (2008) Puntius sarana India JQ667570 Khedkar et al. (2014) Puntius sarana subnasutus HE664122 Vincent (2012) Barbodes Barbodes binotatus Malaysia KT001028 Meganathan et al. (2015) Barbodes binotatus Thailand KP263421 Panprommin and Chanto (2014) Systomus Systomus sarana India JX983454 Khedkar et al. (2014) Systomus sarana India KJ476776 Lal et al. (2014) Systomus orphoides Thailand KP263417 Panprommin and Chanto (2014) Barbus Barbus matthesi Afrika KR153301 Sakharova (2015) Barbus camptacanthus Afrika HM418123 iBOL (2010) Barbus callipterus Afrika HM418124 iBOL (2010) Barbus brazzai Afrika KT193523 Decru et al. (2015) Barbonymus Barbonymus schwanenfeldii Malaysia KT001008 Meganathan et al. (2015) Barbonymus schwanenfeldii Laos JQ346171 Pasco-Viel et al. (2012) Barbonymus altus Afrika KU568767 Van Der Walt et al. (2012) Barbonymus gonionotus Philipina HQ682667 Aquino et al. (2011) Rasbora Rasbora daniconius India JX983478 Khedkar et al. (2012) Danio Danio choprai Myanmar KX852477 Kullander and Noren (2016) Botia China KM610433 Chen et al. (2015)

56 BIODIVERSITAS 20 (1): 54-60, January 2019

RESULT AND DISCUSSION more common than transversion, as found in animal mitochondrial genomes (Briolay et al. 1998). The bias The morphological characters of Puntius cf. binotatus value of transition and transversion ratio (Ts; Tv) is R: in Gunung Tujuh Lake were similar to Punitus binotatus 7.208. P. cf. binotatus from Sumatra showed 71 described by Weber and de Beaufort (1916); Roberts (1989 substitutions which consist of 62 transitions and nine and Kottelat et al. (1993)). The characters that distinguish transversions without insertion and deletion. The COI gene P. cf. binotatus in Gunung Tujuh Lake and P. binotatus is representing 188 amino acids for all samples. Among all from other area are the absence of a black spot on the base of the mutations were observed in P. cf. binotatus of of the anterior dorsal ray and in the part of the body. Sumatra, change has occurred only in one amino acid that According to Vitri et al. (2012), ten morphological is in the 172nd amino acid (I→V). The calculated characters of P. binotatus in West Sumatra were different nucleotide diversity (pi) in the 46 sequences was of significantly (Table 2), and also showed the morphological 0.11608, Notably, for P. cf. binotatus in Sumatra, the variations at a different altitude. nucleotide diversity (pi) was 0.04650, that value is at the The total size of the COI gene is 720 bp and 564 bp of middle level when compared to the other reported species them were successfully amplified. The average of of Puntius (Negi et al. 2017). The value indicated that in nucleotide composition was of A: 27.30%, C: 27.20%, T: Sumatra, there are P. cf. binotatus which vary genetically. 29.40%, and G: 16.10%. The nucleotide composition was Four phylogenetic methods (ME/NJ/ML/MP) were used similar to previous studies (Negi et al. 2017; Roesma et al. to construct the phylogenetic tree (Figure 1) The topologies 2018). The frequency of distributions for the four of ML tree was similar to NJ, ME and MP using 1000 nucleotide bases was significantly different, with non- bootstrap replicates. Two main clusters were obtained, overlapping confidence intervals. The proportion of supported by high bootstrap value of 97/98/95/96 for pyrimidines (T, C) to purines (A, G) deviated from a 50:50 ME/NJ/ML/MP, respectively. The first cluster consists of ratio is well described in the literature for protein-coding five subclusters and the second cluster consists of four genes (Kartavtsev et al. 2009). The bias of the nucleotide subclusters. All the members in the second cluster are the bases reflects the hydrophobic properties of the encoded genus that considered as a synonym to Puntius (Barbus, proteins (Nailor et al. 1996). The A + T content (56.70%) Barbonymus, Puntius, and Systomus). The sequence in present study higher than G + C (43.30%). Those values divergences that separate the first and second cluster was of indicate the characteristic of the mitochondrial DNA of all 14.2-20.4%. When referring to sequence divergence value classes of (Johns and Avise 1998; Kartavtsev et al. reported by Kartavtsev et al. (2007) and Kartavtsev et al. 2007; Kartavtsev et al. 2011; Xiao et al. 2012; Negi et al. (2011), all the member of the first cluster cannot be 2017). considered Puntius. The sequence divergences between P. Of the 564 bp COI gene sequences, 352 bp (62.41%) cf. binotatus in Gunung Tujuh Lake with other P. cf. was conserved sites, and 212 bp (37.59%) was the variable binotatus in Sumatra were of 3.1-7.4%. site without insertion or deletion. In the total of 212 In the first subcluster of the topology, P. cf. Binotatus variable sites, 183 bp (86.32%) are parsimony informative Gunung Tujuh Lake is in the same cluster with P. cf. site, and 29 bp (13.68%) are singleton site. In total, there binotatus from Diatas lakes, Batang Lembang, Batang were 337 mutations in all sequences, the transitions are Gumanti, Muara Pingai rivers, and P. banksi from Malaysia.

Table 2. Morphometric measurements of Puntius cf. binotatus in Gunung Tujuh Lake, Kerinci, Jambi, Indonesia

Characteristics Code Range (mm) n=50 %Standar Length (SL) Max Min Mean ± SD Max Min Mean ± SD Total length TL 80.54 49.2 61.90 ± 7.12 147.24 105.56 129.36± 5.28 Standard length SL 62.83 37.7 47.93± 5.96 - - - Head length HL 18.22 11.2 14.08± 1.54 40.02 25.26 29.51± 2.35 Head Depth HD 12.16 6.95 9.73± 1.10 30.30 14.44 20.42± 2.26 Head Width HW 10.52 4.87 7.11± 1.05 20.55 11.95 14.88± 1.69 Eye Diameter ED 4.15 2.07 2.89± 0.34 8.23 4.03 6.09± 0.84 Interorbital width IW 5.96 3.43 4.76± 0.58 14.95 7.69 9.99± 1.20 Mouth Length ML 27.1 1.42 2.67± 3.54 53.01 2.95 5.55± 6.90 Interorbital length IOL 14.6 8.92 10.94± 1.27 25.68 19.01 22.89± 1.43 Snout of length SNL 23.39 2.46 6.24± 4.15 54.06 6.02 13.20± 9.44 Body Depth BD 17.08 4.99 9.67± 3.72 33.64 10.67 20.14± 7.13 Body Width BW 20.43 4.44 10.81± 3.98 34.94 10.85 22.60± 8.00 Caudal Peduncle Length CPL 11.39 6.24 8.78± 1.04 29.13 13.42 18.45± 2.38 Dorsal Base Length DBL 8.68 4.12 6.74±0.95 18.40 10.06 14.13± 1.77 Dorsal Fin Height DFH 13.96 7.27 9.96± 1.46 32.32 15.62 20.92± 3.01 Ventral Fin Length VFL 10.79 5.1 7.97± 1.21 21.30 12.75 16.67± 1.95 Anal Fin Length AFL 9.42 4.97 7.03± 0.92 19.03 10.33 14.77± 1.87

ROESMA et al. – Taxonomy confirmation of Puntius cf. binotatus 57

The sequence divergences between P. cf. binotatus Gunung number of JN646096 in the fifth subcluster of the first Tujuh Lake and other members in that first subcluster are cluster. Based on the values of Ward et al. (2005), 3.1-4.1%, supported with the highly bootstrap value Kartavtsev et al. (2009), Kartavtsev (2011), and Kartavtsev (99/99/96/98 for ME/NJ/ML/MP, respectively). Sequence (2013), the range of 11.6-13% value shows that all of the divergences between P. cf. binotatus Gunung Tujuh Lake four subclusters differ at the species level to P. binotatus and P. banksi from Malaysia was 4.1%. Referring to the Malaysia with the accession number of JN646096. values reported by Ward et al. (2005), Kartavtsev et al. Puntius binotatus from Malaysia with the accession (2009), Kartavtsev (2011), and Kartavtsev (2013), this number JN646096 occupies the position at the cladogram value places P. cf. binotatus Gunung Tujuh Lake is at the that suggests it as the common ancestor to all of Puntius in sub-species level. Based on Cyt b gene, Karlina et al. Sumatra. In NCBI data Genbank, P. binotatus Malaysia (2016) reported that the sequence divergences between P. with the accession number JN646096 and P. binotatus cf. binotatus Gunung Tujuh Lake and others population (P. Lampung with the accession number JQ665834 listed as cf. binotatus from Diatas lakes, Batang Lembang, Batang Barbodes binotatus while in the IUCN Red List the Gumanti, Muara Pingai rivers and P. banksi from Barbodes binotatus stated as the accepted scientific/valid Malaysia) were 6.6-8.1% which positions it as a name of Puntius binotatus (Eschmeyer 2015; Jenkins et al. subspecies. Therefore both types of the genes provide the 2015; IUCN 2018). In addition, B. binotatus Thailand same conclusion that P. cf. binotatus Gunung Tujuh Lake (KP263421), B. binotatus Malaysia (KT001028), and B. is at the level of sub-species. binotatus Lampung (JQ665834) cannot be classified as P. Puntius cf. binotatus Gunung Tujuh Lake is a sister binotatus. Hence, all member of the first cluster is taxon of P. cf. binotatus from other populations in the first considered as the genus of Barbodes. subcluster. This closed relationship may be because of two The genetic distance between Puntius banksi Malaysia group of fish live in the same geographical and ecological (JF781235) and other members at same branch (P. cf. conditions. All populations of P. cf. binotatus in the first binotatus Batang Gumanti River, P. cf. binotatus Diatas subcluster are located at the highland over 1.500 meters Lake, P. cf. binotatus Muaro Pingai River, and P. cf. above sea level in the eastern part of Bukit Barisan binotatus Batang Lembang River) is ranged between 0.9% Mountain range (Roesma et al. 2018). Bukit Barisan to1.3%, thus this group is identified as B. banksi. This Mountain range is the row of the mountain that spreading results of this study implied that there are four subspecies out from northern to southern of Sumatra island more or of B. banksi in Sumatra. P. cf. binotatus Gunung Tujuh less 1650 km in length. It is estimated that these conditions Lake is proposing as B. banksi gunungtujuh. cause local adaptations to bring their current genetic Studies by those of Kartavtsev and Lee (2006); variation. Negi et al. (2017) reported that P. sophree from Kartavtsev et al. (2007); Kartavtsev et al. (2009); the distinct region with the most homogeneous Kartavtsev (2011); Kartavtsev et al. (2014) reported that environment has low genetic diversity compared the the range of sequence divergences values of COI gene populations from the small geographical region with a between P. cf. binotatus Gunung Tujuh and other Sumatra heterogeneous environment. P. cf. binotatus was of 3.1-7.6%, indicating differences at The sequence divergences between Puntius cf. the subspecies level. However, a study by Karlina et al. binotatus Gunung Tujuh Lake and other members in the (2016) using the Cyt b gene suggested a sequence same subcluster with the member of the second subcluster divergence value of 6.6-11.2% indicating differences at (Barbodes binotatus Thailand with the accession number of different species levels. According to (Hebert et al. 2003a; KP263421 and Barbodes binotatus Malaysia with the Hebert et al. 2003b) the COI gene is more representative to accession number of KT001028) are 3.7-4.5%. be used to distinguish species. This support P. cf. binotatus Consequently, P. cf. binotatus Gunung Tujuh Lake and Gunung Tujuh as a subspecies (B. banksi gunungtujuh). other members in the first subcluster can be grouped in The second cluster consists of four sub-clusters consisted Barbodes because of the sequence divergences value do not of genera that synonym of Puntius (Barbus, Barbodes, support them to differ at genus levels. Besides the position Systomus, and Barbonymus). This group is supported by of the branch in the tree, this provision also refers to Ward high bootstrap values (98/97/95/97 for ME/NJ/ML/MP et al. (2005), Kartavtsev et al. (2009), Kartavtsev (2011), respectively). The sequence divergences between genera in and Kartavtsev (2013). the second cluster are 10.7-24.1% and with the first cluster The sequence divergences between the first and second is 13.7-21.1%. These values show that they are different subclusters in the first cluster with the member of the third genus but the same family. At the second cluster of each subcluster (Puntius cf. binotatus Batang Kuranji River, P. genus that is considered a synonym of Puntius clustered on cf. binotatus Batang Tarok River, and P. cf. binotatus different sub-clusters, there are overlapping of placements Batang Tarok River) are 6.3-7.0%. That value indicated in the Systomus and Puntius groups in the third subclasses. that they are different at the subspecies level. Furthermore, In the IUCN Red List, Fish Base and Catalog of life the sequence divergences between the first, second, third (Dahanukar 2010; Froese and Pauly 2018; IUCN 2018) the subclusters in the first cluster with the member of the S. sarana and S. orphoides stated as the accepted fourth subcluster are of 6.7-8.5%. Those values also scientific/valid name of P. sarana and P. orphoides. indicated that they are different the subspecies. All of the Therefore, the members of the third subcluster can be four subclusters have the sequence divergences value of specified as Systomus group. 11.6-13% to P. binotatus Malaysia with the accession

58 BIODIVERSITAS 20 (1): 54-60, January 2019

Figure 1. The ML phylogenetic tree of COI gene with bootstrapping of 1000 replicates (ME/ML/NJ/MP)

It is concluded that P. cf binotatus group in Sumatra first cluster (Barbodes) and second cluster (Barbus, can be considered as a Barbodes banksi that further Puntius, Systomus, and Barbonymus). Moreover, the results consisted of four subspecies, one of them is P. cf. binotatus of the analysis do not support the placement of Barbodes, Gunung Tujuh Lake named as Barbodes banksi Barbus, Systomus, and Barbonymus as synonyms of the gunungtujuh. In the present phylogenetic tree, there is a Puntius. clear separation or grouping of each different genus in the

ROESMA et al. – Taxonomy confirmation of Puntius cf. binotatus 59

ACKNOWLEDGMENTS Haryono, Tjakrawidjaja AH. 2006. Morphological study for identification improvement of tambra fish (Tor Spp.: Cyprinidae) from Indonesia. Biodiversitas 7 (1) : 59-62. The Authors thanks the Management of the Ministry of Hebert PDN, Cywinska A, Ball SL, de Waard JR. 2003a. Biological Research, Technology and Higher Education whose identifications through DNA barcodes. Proc R Soc London B 270: support the funding of the research to Andalas University. 313-321. Our thanks also expressed to Biology Department of Hebert PDN, Ratnasingham S, de Waard JR 2003b. Barcoding Animal Andalas University for field and Laboratory work permit. Life: Cytochrome c Oxidase Subunit 1 Divergences among Closely Related Species Proc R Soc London B (Suppl.) 270: (S96–S99). We also would like to thank our students who helped us in Hebert PDN, Gregory TR. 2005. TR: The promise of DNA barcoding for sample collection and laboratory works in Genetic and taxonomy. Syst Biol 54 (5): 852-859. Biomolecular Laboratory, Faculty of Mathematics and International Barcode of Life (iBOL). 2010. iBOL Data Release. Sciences, Andalas University, Padang, Indonesia. www.gbif.org/search?q=Barbus IUCN. 2018. The IUCN Red List of Threatened Species. Version 2018. ISSN 2307-8235 Jenkins A, Kullander FF, Tan HH. 2015. Barbodes binotatus. The IUCN REFERENCES Red List of Threatened Species 2015: e.T169538A70031333. DOI: 10.2305/IUCN.UK.2015-1.RLTS.T169538A70031333.en. John L, Peter R, Gopalakrishnan A. 2013. Population structure of Aquino LM, Tango JM, Canoy RJ, Fontanilla IK, Basiao ZU, Ong PS, Denison’s barb, Puntius denisonii (Pisces: Cyprinidae): a species Quilang JP. 2011. DNA barcoding of fishes of Laguna de Bay, complex endemic to the Western Ghats of India. J Phylogen Evol Biol Philippines. Mitochond DNA 22 (4): 143-153. 1: 106. Briolay J, Galtier N, Brito RM, Bouvet, Y. 1998. Molecular Phylogeny of Johns GC, Avise JC. 1998. A comparative summary of genetic distance Cyprinidae Inferred from cytochrome bDNA Sequences. Molecular from mitochondrial cytochrome b gene. Mol Biol Evol Phylogenet Evol 9 (1): 100-108. 15: 1481-1490. Burland TG. 2000. DNASTAR’s Lasergene Sequence Analysis Software. Karlina W, Roesma DI, Tjong DH. 2016. Phylogenetic study of Puntius Methods Mol Biol 132: 71-91. cf. binotatus fish from Gunung Tujuh Lake in Sumatera Based on Cailliet GM, Love MS, Ebeling AW. 1986. Fishes: A Field and Cytochrome b Gene. J Entomol Zool Stud 4 (2): 538-540. Laboratory Manual on Their Structure, Identification, and Natural Kartavtsev YP. 2013. Sequence Diversity at Cyt-b and Co-1 mtDNA History. Waveland Press, Inc., Long Grove, IL. Genes in Animal Taxa Proved Neo-darwinism. J Phylogenet Evol Champasri T, Rapley R, Duangjinda M, Suksri A. 2007. A Morphological Biol 1(4):1-5. Identification in Fish of the Genus Puntius Hamilton 1822 Kartavtsev YP, Lee JS. 2006. Analysis of nucleotide diversity at the (: Cyprinidae) of Some Wetlands in Northeast cytochrome b and Cytochrome Oxidase-I genes at the population, Thailand. Pak J Biol Sci 10 (24): 4383-4390. species, and genus levels. Russ J Genet 42 (4): 341-362 Chen W, Ma X, Shen Y, Mao Y, He S. 2015. The fish diversity in the Kartavtsev YP, Park TJ, Vinnikov KA, Ivankov VN, Sharina SN, Lee JS. upper reaches of the Salween River, Nujiang River, revealed by DNA 2007. Cytochrome b (Cyt-b) gene sequences analysis in six flatfish barcoding. Sci Rep 5: 17437. species (Pisces, Pleuronectidae), with phylogenetic and taxonomic Clare EB, Lim BK, Engstrom MD, Eger JL, Hebert PDN. 2006. DNA insights. Mar Biol 152 (4): 757-773. barcoding of Neotropical Bats: species identification and discovery Kartavtsev YP, Sharina SN, Goto T, Balanov AA, Hanzawa, N. 2009. within Guyana. Mol Ecol Notes 7: 184-190. Sequence diversity at Cytochrome Oxidase-I (Co-1) gene among Collins RA, Yi Y, Meier R, Armstrong K. 2011. Barbs, barcodes, and sculpins (Scorpaeniformes, Cottidae) and some other scorpionfish of biosecurity: identifying cyprinid fishes in the ornamental trade. Russia Far East with phylogenetic and taxonomic insights. Genes National University of Singapore, Singapore. Genom 31 (2): 191-205. Collins RA, Armstrong KF, Meier R, Yi Y, Brown SDJ, Cruickshank RH, Kartavtsev YP. 2011. Divergence at Cyt-b and Co-1 mtDNA genes on Keeling S, Johnston C. 2012. Barcoding and border biosecurity: different taxonomic levels and genetics of speciation in . Identifying cyprinid fishes in the aquarium trade. PLoS ONE 7 (1): Mitochond DNA 3: 55-65. e28381. DOI: 10.1371/journal.pone.0028381. Kartavtsev YP, Rozhkovan KV, Masalkova NA. 2014. Phylogeny based Dahanukar N. 2010. Systomus sarana. The IUCN Red List of Threatened on two mtDNA genes (COI, Cytb) among Sculpins (Scorpaeniformes, Species e.T166567A6237905. DOI: 10.2305/IUCN.UK.20104.RLTS. Cottidae) and some other scorpionfish in the Russian Far East. T166567A6237905.en. Mitochond DNA 27: 2225-2240. Decru E, Moelants T, De Gelas K, Vreven E, Verheyen E, Snoeks J. 2015. Khedkar GD, Jamdade R, Naik S, David L, Haymer D. 2014. DNA Taxonomic challenges in freshwater fishes: a mismatch between barcodes for the fishes of the Narmada, One of India's Longest morphology and DNA barcoding in fish of the north-eastern part of Rivers. PLoS ONE 9 (7): E101460. DOI: 10.1371/journal the Congo basin. Mol Ecol Resour 16 (1): 342-352. .pone.0101460. Eschmeyer WN. (ed.). 2015. Catalog of Fishes. Updated 2 October 2018. Kottelat M. 2013. The fishes of the inland waters of Southeast Asia: a www.calacademy.org/scientists/projects/catalog-of-fishe catalogue and core bibliography of the fishes known to occur in Frezal L, Leblois R. 2008. Four years of DNA barcoding: current freshwaters, mangroves and estuaries. Raffles Bull Zool 27:1-663. advances and prospects. Infection, genetics, and evolution. J Mol Kottelat M, Whitten AJ., Kartikasari SN, Wiejoatmodjo S. 1993. Epidemiol Evol Genet Infect Dis 8: 727-736. Freshwater Fishes of Western Indonesia and Sulawesi. Periplus Eds. Froese R, Pauly D. (eds). 2018. FishBase (version Feb 2018). In: Roskov (HK) Ltd., Singapore. Y, Ower G, Orrell T, Nicolson D, Bailly N, Kirk PM, Bourgoin T, Kullander SO, Noren M. 2016. Danio htamanthinus (Teleostei: DeWalt RE, Decock W, Nieukerken EV, Zarucchi J, Penev L, eds. Cyprinidae), a new species of miniature cyprinid fish from the Species 2000 and ITIS Catalogue of Life, 30th October 2018. Digital Chindwin River in Myanmar. Zootaxa 4178 (4): 535-546. resource at www.catalogueoflife.org/col. Species 2000: Naturalis, Lakra WS, Verma MS, Goswami M. 2008. DNA Barcoding of Indian Leiden, the Netherlands. Freshwater Fishes. National Bureau of Fish Conservation Division, Hajibabaei M, Singer GAC, Clare EL, Hebert PDN. 2007. Design and NBFGR,Uttar Pradesh, India. applicability of DNA arrays and DNA barcodes in biodiversity Lakra WS, Goswami M, Yadav K. 2010. Development and monitoring. BMC Biol 5: 1-15. characterization of cell lines from Denison's barb. National Bureau of Hall, TA. 1999. BioEdit: A user-friendly biological sequence alignment Fish Conservation Division, NBFGR,Uttar Pradesh, India. editor and analysis program for Windows 95/98/NT. Nucleic Acid Lal KK, Mohindra V, Singh RK, Dhawan S, Chandra S, Gupta BK, Symp Ser 41: 95-98. Dwivedi AK, Jena JK. 2014. Molecular characterization of Indian Handika HA, Mursyid A, Chornelia A, Akbar MA, Karlina W, Roesma freshwater Cyprinids using cytochrome C Oxidase I sequences. DI. 2014. Diversitas Ikan, Burung (Khusus Famili Timalidae) dan National Bureau of Fish Conservation Division, NBFGR,Uttar Mamalia kecil di Gunung Tujuh dan Masurai. Laporan Penelitian. Pradesh, India. Universitas Andalas, Padang. [Indonesian]

60 BIODIVERSITAS 20 (1): 54-60, January 2019

Meganathan P, Austin CM, Tam SM, Chew PC, Siow R, Rashid AZ, Sakharova H. 2015. Barbin cyprinids of the Kwilu River (Democratic Song BK. 2015. An application of DNA barcoding to the Malaysian Republic of Congo) with a description of a new species of freshwater fish fauna: mtDNA COI sequences reveal novel . Democratic Republic of Congo, Afrika. haplotypes, cryptic species, and field-based misidentification. Monash Singh NS, Behera BK, Sharma AP. 2013. Population Structure of Puntius University Malaysia, Selangor, Malaysia. sophore Inferred from Variation in Mitochondrial DNA Sequences. Mohsin AKM, Ambak, MA. 1983. Freshwater fishes of Peninsular International J Research in Fisheries and Aquaculture 3 (3): 112-115. Malaysia. University Pertanian Malaysia, Malaysia. Smith MA, Rodriguez JJ, Whitfield JB, Deans AR, Janzen DH. 2008. Nailor GJ, Collins TM, Brown WM. 1996. Hydrophobicity and Extreme diversity of tropical parasitoid wasps exposed by iterative phylogeny. Nature 373: 565-566. integration of natural history, DNA barcoding, morphology, and Negi RK, Joshi BD, Johnson JA, De R, Goyal SP. 2017. Phylogeography collections. Proc Natl Acad Sci USA 105: 12359-12364. of freshwater fish Puntius sophore in India. Mitochond DNA 29 : Song LM, Munian K, Abd Rashid Z, Bhassu, S. 2013. Characterization of 256-265. Asian Snakehead Murrel Channa striata (Channidae) in Malaysia: An Panprommin D, Chanto W. 2014. Fish species diversity and identification Insight into Molecular Data and Morphological Approach. The using DNA barcode in Than. Sawan Waterfall, Doi Phu Nang Scientific World Journal. DOI: 10.1155/2013/917506. national park, Phayao province. Phayao University, Phayao, Thailand. Stoeckle M. 2003. Taxonomy, DNA and the barcode of life. Bio Sci 53: Pasco-Viel E, Veran M, Viriot L. 2012. Bleeker was right: Revision of the 796-797. genus Cyclocheilichthys (Bleeker 1859) and resurrection of the genus Taki Y, Katsuyama A, Urushido T. 1978. Comparative Morphology and Anematichthys (Bleeker 1859), based on morphological and Interspecific Relationships Of The Cyprinid Genus Puntius. Japanese molecular data of Southeast Asian Cyprininae (Teleostei, J Ichthyol 25: 1-8. Cypriniformes). Zootaxa 3586: 41-54. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. 2013. MEGA 6: Rainboth, WJ. 1996. Fishes of the Cambodian FAO Species molecular evolutionary genetics analysis version 6.0. Mol Biol Evol Identification Field Guide of Fishery Purpose. Rome: Food and 30: 2725-2729. Agriculture Organization: 1-265. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. Roberts TR. 1989. The Freshwater Fishes of Western Borneo (Kalimantan The Clustal X windows interface flexible strategies for multiple Barat, Indonesia). California Academy of Sciences 14: 1-210. sequence alignment aided by quality analysis tools. Nucleic Acids Roesma DI. 2011. Diversitas Species dan Kekerabatan Genetik Ikan-Ikan Research 24: 4876-4882. Cyprinidae di Danau-Danau dan Sungai-Sungai di Sekitarnya Di Van Der Walt KA, Makinen T, Weyl O, Collins R, Swartz E. 2012. DNA Kawasan Sumatera Barat. Disertasi. Universitas Andalas, Padang. barcoding of ornamental freshwater fish from South Africa. South (Species Diversity and Genetic Relationship of Cyprinidae in Lakes African Institute for Aqutic Biodiversity, Eastern Cape, South Africa. and the Adjoining Rivers in West Sumatra. Dissertation. Andalas Vincent M. 2012. Visual and olfactory perception in the reproduction of a University, Padang). [Indonesian] cyprinid. Fish P. parrah Day. [Thesis]. Calicut University, Roesma DI, Chornelia A, Mursyd A, Kamsi M. 2016. Short Mallapuram. [Indian] Communication: Fish diversity of the Batang Toru River System, Vitri DK, Roesma DI, Syaifullah. 2012. (Morphological Analysis of P. South Tapanuli, North Sumatra. Biodiversitas. 17 (2): 628-634 binotatus Valenciennes 1842 (Pisces: Cyprinidae) from several Roesma DI, Djong HT, Munir W, Agesi AV, Chornelia A. 2017. Genetic locations in West Sumatra. Jurnal Biologi Universitas Andalas 1 (2): diversity of Tor douronensis (Pisces: Cyprinidae) in West Sumatra, 139-143. Indonesia Biodiversitas 18 (3): 1018-1025. Ward RD, Zemlak TS, Innes BH, Last PR, Hebert PDN. 2005. DNA Roesma DI, Djong HT, Munir W, Aidil DR. 2018. New Record Species of barcoding Australia’s fish species. Philos Trans R Soc B 360: 1847- Puntius (Pisces: Cyprinidae) from West Sumatra Based on 1857. Cytochrome Oxidase I Gene. International Journal on Advanced Weber MG, de Beaufort LF. 1916. Fishes of the Indo-Australian Science Engineering Information Technol 8 (1): 250-256. Archipelago Vol. III. E.J. Brill, Leiden. Rozas J, Sanchez-Del Barrio JC, Messeguer X, Rozas R. 2003. DNA SP, Wibowo A, Farajalah A, Husnah H. 2012. DNA barcoding of freshwater DNA polymorphism analyses by coalescent and other methods. fish species of Manna River (Bengkulu) and Semanka River Bioinformatics 19: 2496-2497. (Lampung). Indonesia Fish Res J 19 (1): 9-17. Sade A, Biun, H. 2012.The Ichthyofauna of Maliau Basin Buffer Zone at Xiao B, Chen AH, Zhang YY, Jiang GF, Hu CC, Zhu CD. 2012. Maliau Basin. Conservation Area, Sabah, Malaysia. J Trop Biol Complete mitochondrial genomes of two cockroaches, Blattella Conserv 9 (1): 105-113. germanica and Periplaneta americana, and the phylogenetic position of termites. Curr Genet 58 (2) : 65-77.